Integrin-mediated migration of neutrophils to infected tissue sites is vital for pathogen clearance and therefore host survival. Although β₂ integrins have been shown to mediate neutrophil transendothelial migration during systemic and local inflammation, relatively little information is available regarding neutrophil migration in sepsis beyond the endothelial cell layer. In this study, we report that integrin α₃β₁ (VLA-3; CD49c/CD29) is dramatically upregulated on neutrophils isolated from both human septic patients and in mouse models of sepsis. Compared with the α₃β₁^low granulocytes, α₃β₁^high cells from septic animals displayed hyperinflammatory phenotypes. Administration of a α₃β₁ blocking peptide and conditional deletion of α₃ in granulocytes significantly reduced the number of extravasating neutrophils and improved survival in septic mice. In addition, expression of α₃β₁ on neutrophils was associated with Toll-like receptor–induced inflammatory responses and cytokine productions. Thus, our results show that α₃β₁ is a novel marker of tissue homing and hyperresponsive neutrophil subtypes in sepsis, and blocking of α₃β₁ may represent a new therapeutic approach in sepsis treatment.